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首页> 外文期刊>Photodiagnosis and Photodynamic Therapy >Autofluorescence spectroscopy for NADH and flavoproteins redox state monitoring in the isolated rat heart subjected to ischemia-reperfusion
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Autofluorescence spectroscopy for NADH and flavoproteins redox state monitoring in the isolated rat heart subjected to ischemia-reperfusion

机译：自体荧光光谱法检测离体大鼠心脏缺血再灌注后NADH和黄素蛋白的氧化还原状态

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摘要

Background: Reduction of myocardial ischemia-reperfusion injury in the patients undergoing cardiac surgery under cardiopulmonary bypass represents an important goal. Intraoperative monitoring of myocardial metabolic state using continuous registration of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) fluorescence might contribute to the solution of the problem. The successful application of fluorescent spectroscopy in the clinical field requires additional refinement of the technique, particularly using excitation of both NADH and FAD with different wavelengths. Methods: The experiments were performed on the isolated Langendorff-perfused rat hearts (n = 28) subjected to either regional or global ischemia-reperfusion. Two principles of NADH and FAD autofluorescence (AF) measurement were used for ischemia monitoring: analysis of photographs and videos obtained with multispectral organoscopy technique allowing the assessment of both spatial and temporal characteristics of the process (n = 16); continuous registration of tissue redox state in a representative area of the heart by application of local spectroscopy, assisted by fiber optic spectrometer (n = 12). Results: It was found that regional myocardial ischemia resulted in a rapid, substantial increase in the intensity of NADH AF excited at 360 nm in the ischemic versus non-ischemic area of the heart. The same result was obtained when the heart was made globally ischemic, while the restoration of perfusate flow completely reversed the increase in NADH AF. During the transition from ischemia to reperfusion, the spatial heterogeneity of myocardial AF was noted on video recordings, probably reflecting the microheterogeneity of myocardial blood flow. Local spectroscopy studies demonstrated opposite changes in the NADH and FAD AF during ischemia. Using both methodological approaches, we found that repetitive brief episodes of global myocar-dial ischemia resulted in progressive decrease in the magnitude of AF elevation, which might point to preconditioning effect. Conclusions: The application of multispectral fluorescent organoscopy offers the advantage of monitoring myocardial redox state at the level of the entire heart. Local spectroscopy is characterized by better precision and, in addition, provides the unique opportunity to measure AF in different parts of the spectrum. AF measurements are non-invasive, rapid, and technically easy to perform. For future clinical applications, it might be recommended to combine the measurement of redox state of both NADH and FAD, using excitation wavelength and emission filter optimal for each fluorophore.
机译：背景：减少体外循环下进行心脏手术的患者的心肌缺血-再灌注损伤是一个重要的目标。使用烟酰胺腺嘌呤二核苷酸（NADH）和黄素腺嘌呤二核苷酸（FAD）荧光的连续配准术中监测心肌代谢状态可能有助于解决该问题。荧光光谱法在临床领域的成功应用需要对该技术进行进一步的改进，尤其是使用不同波长的NADH和FAD进行激发。方法：实验是在离体的Langendorff灌注大鼠心脏（n = 28）上进行的，该大鼠心脏经历了局部或整体缺血再灌注。 NADH和FAD自发荧光（AF）测量的两个原理用于缺血监测：分析通过多光谱器官镜技术获得的照片和视频，从而可以评估过程的时空特性（n = 16）；通过局部光谱在光纤光谱仪（n = 12）的辅助下连续记录心脏代表区域中组织的氧化还原状态。结果：发现局部心肌缺血导致心脏缺血区域和非缺血区域在360 nm激发的NADH AF强度迅速，显着增加。当心脏整体缺血时，获得相同的结果，而灌注液流量的恢复完全逆转了NADH AF的增加。在从缺血到再灌注的过渡过程中，视频记录中记录了心肌AF的空间异质性，这可能反映了心肌血流的微异质性。局部光谱研究表明，缺血期间NADH和FAD AF的变化相反。使用这两种方法学方法，我们发现整体性心肌缺血的反复短暂发作导致房颤升高的幅度逐渐降低，这可能预示了预处理的作用。结论：多光谱荧光器官镜检查的应用提供了在整个心脏水平上监测心肌氧化还原状态的优势。局部光谱的特点是精度更高，此外，它还提供了测量光谱不同部分的AF的独特机会。 AF测量是非侵入性的，快速的并且在技术上易于执行。对于将来的临床应用，可能建议使用对于每种荧光团最佳的激发波长和发射滤光片，将NADH和FAD的氧化还原状态的测量结果结合起来。

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  • 来源
    《Photodiagnosis and Photodynamic Therapy》 |2014年第3期|400-408|共9页
  • 作者

    Garry Papayan; Nickolay Petrishchev; Michael Galagudza;

  • 作者单位

    Center of Laser Medicine, Department of Pathophysiology, First I.P. Pavlov Federal Medical University of St. Petersburg, Lev Tolstoy Str. 6/8, 197022 St. Petersburg, Russian Federation,Institute of Experimental Medicine, Federal Almazov Medical Research Centre, Akkuratova Str. 2, 197341 St. Petersburg, Russian Federation;

    Center of Laser Medicine, Department of Pathophysiology, First I.P. Pavlov Federal Medical University of St. Petersburg, Lev Tolstoy Str. 6/8, 197022 St. Petersburg, Russian Federation,Institute of Experimental Medicine, Federal Almazov Medical Research Centre, Akkuratova Str. 2, 197341 St. Petersburg, Russian Federation;

    Center of Laser Medicine, Department of Pathophysiology, First I.P. Pavlov Federal Medical University of St. Petersburg, Lev Tolstoy Str. 6/8, 197022 St. Petersburg, Russian Federation,Institute of Experimental Medicine, Federal Almazov Medical Research Centre, Akkuratova Str. 2, 197341 St. Petersburg, Russian Federation;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    Photodiagnosis; Autofluorescence; Spectroscopy; Nicotinamide adenine dinucleotide; Heart; Ischemia;

    机译：光诊断;自发荧光;光谱学;烟酰胺腺嘌呤二核苷酸;心;缺血;

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